Purpose: The genomic characteristics of uterine sarcomas have not been fully elucidated. This study aimed to explore the genomic landscape of the uterine sarcomas (USs). Materials and Methods: Comprehensive genomic analysis through RNA-sequencing was conducted. Gene fusion, differentially expressed genes (DEGs), signaling pathway enrichment, immune cell infiltration, and prognosis were analyzed. A deep learning model was constructed to predict the survival of US patients. Results: A total of 71 US samples were examined, including 47 endometrial stromal sarcomas (ESS), 18 uterine leiomyosarcomas (uLMS), three adenosarcomas, two carcinosarcomas, and one uterine tumor resembling an ovarian sex-cord tumor. ESS (including high-grade ESS [HGESS] and low-grade ESS [LGESS]) and uLMS showed distinct gene fusion signatures; a novel gene fusion site, MRPS18A–PDC-AS1 could be a potential diagnostic marker for the pathology differential diagnosis of uLMS and ESS; 797 and 477 uterine sarcoma DEGs (uDEGs) were identified in the ESS vs. uLMS and HGESS vs. LGESS groups, respectively. The uDEGs were enriched in multiple pathways. Fifteen genes including LAMB4 were confirmed with prognostic value in USs; immune infiltration analysis revealed the prognositic value of myeloid dendritic cells, plasmacytoid dendritic cells, natural killer cells, macrophage M1, monocytes and hematopoietic stem cells in USs; the deep learning model named Max-Mean Non-Local multi-instance learning (MMN-MIL) showed satisfactory performance in predicting the survival of US patients, with the area under the receiver operating curve curve reached 0.909 and accuracy achieved 0.804. Conclusion USs harbored distinct gene fusion characteristics and gene expression features between HGESS, LGESS, and uLMS. The MMN-MIL model could effectively predict the survival of US patients.

Objective To investigate the role and mechanism of the zinc finger protein Kruppel-like transcription factor 9 (KLF9) in the stimulation of type I interferon expression induced by herpes simplex virus type 1 (HSV-1) in macrophages. Methods Agarose Gel electrophoresis, quantitative real-time PCR (qRT-PCR) and western blot analyses were employed to detect the KLF9 relative expression in bone marrow-derived macrophages (BMDMs) from Klf9^-/- (gKO) mice and wild-type (WT) mice. RNA-seq analysis was utilized to identify the potential targeted genes upon HSV-1 stimulation in BMDMs. ELISA was used to measure the potent of IFN-β in the supernatant of BMDMs derived from gKO and WT mice after HSV-1 stimulation. qRT-PCR analysis was employed to further confirm the changes of Ifnb1 and interferon-stimulated gene (ISG) such as interferon-induced protein with tetratricopeptide repeats 1 (Ifit1), interferon-stimulated exonuclease gene 20 (Isg20), cholesterol 25-hydroxylase (Ch25h) and 2'-5' oligoadenylate synthetase-like 1 (Oasl1). Western blot was used to detect the expression of phosphorylated interferon regulatory factor-3 (p-IRF3), IRF3, phosphorylated interferon regulatory factor-7 (p-IRF7), IRF7, phosphorylated nuclear factor-kappa B p65 (p-NF-κB p65) and NF-κB p65. CUT-Tag and ChIP-qPCR assay were utilized to confirm the binding region of KLF9 in Ifnb1. Results The KLF9 expression was significantly decreased in BMDMs from gKO mice compared with that from WT mice. The RNA-seq analysis showed that Klf9 deletion in BMDMs resulted in an impaired type I interferon signaling pathway. The qRT-PCR analysis revealed that Klf9 deletion in BMDMs led to a significant decrease of Ifnb1 and ISG such as Ifit1, Ch25h and Oasl1 except Isg20. Moreover, ELISA revealed that Klf9 knockout in BMDMs resulted in a significant decrease of IFN-β secreted from BMDMs. Mechanistically, KLF9 directly binds to the promoter of Ifnb1. Conclusion KLF9 is essential for macrophages to resist HSV-1 infection.
Animals ; Kruppel-Like Transcription Factors/physiology* ; Interferon-beta/metabolism* ; Macrophages/virology* ; Mice ; Herpesvirus 1, Human/physiology* ; Mice, Knockout ; Signal Transduction ; Mice, Inbred C57BL ; Interferon Regulatory Factor-3/genetics* ; Interferon Regulatory Factor-7/genetics* ; Gene Expression Regulation

Purpose: The genomic characteristics of uterine sarcomas have not been fully elucidated. This study aimed to explore the genomic landscape of the uterine sarcomas (USs). Materials and Methods: Comprehensive genomic analysis through RNA-sequencing was conducted. Gene fusion, differentially expressed genes (DEGs), signaling pathway enrichment, immune cell infiltration, and prognosis were analyzed. A deep learning model was constructed to predict the survival of US patients. Results: A total of 71 US samples were examined, including 47 endometrial stromal sarcomas (ESS), 18 uterine leiomyosarcomas (uLMS), three adenosarcomas, two carcinosarcomas, and one uterine tumor resembling an ovarian sex-cord tumor. ESS (including high-grade ESS [HGESS] and low-grade ESS [LGESS]) and uLMS showed distinct gene fusion signatures; a novel gene fusion site, MRPS18A–PDC-AS1 could be a potential diagnostic marker for the pathology differential diagnosis of uLMS and ESS; 797 and 477 uterine sarcoma DEGs (uDEGs) were identified in the ESS vs. uLMS and HGESS vs. LGESS groups, respectively. The uDEGs were enriched in multiple pathways. Fifteen genes including LAMB4 were confirmed with prognostic value in USs; immune infiltration analysis revealed the prognositic value of myeloid dendritic cells, plasmacytoid dendritic cells, natural killer cells, macrophage M1, monocytes and hematopoietic stem cells in USs; the deep learning model named Max-Mean Non-Local multi-instance learning (MMN-MIL) showed satisfactory performance in predicting the survival of US patients, with the area under the receiver operating curve curve reached 0.909 and accuracy achieved 0.804. Conclusion USs harbored distinct gene fusion characteristics and gene expression features between HGESS, LGESS, and uLMS. The MMN-MIL model could effectively predict the survival of US patients.

Purpose: The genomic characteristics of uterine sarcomas have not been fully elucidated. This study aimed to explore the genomic landscape of the uterine sarcomas (USs). Materials and Methods: Comprehensive genomic analysis through RNA-sequencing was conducted. Gene fusion, differentially expressed genes (DEGs), signaling pathway enrichment, immune cell infiltration, and prognosis were analyzed. A deep learning model was constructed to predict the survival of US patients. Results: A total of 71 US samples were examined, including 47 endometrial stromal sarcomas (ESS), 18 uterine leiomyosarcomas (uLMS), three adenosarcomas, two carcinosarcomas, and one uterine tumor resembling an ovarian sex-cord tumor. ESS (including high-grade ESS [HGESS] and low-grade ESS [LGESS]) and uLMS showed distinct gene fusion signatures; a novel gene fusion site, MRPS18A–PDC-AS1 could be a potential diagnostic marker for the pathology differential diagnosis of uLMS and ESS; 797 and 477 uterine sarcoma DEGs (uDEGs) were identified in the ESS vs. uLMS and HGESS vs. LGESS groups, respectively. The uDEGs were enriched in multiple pathways. Fifteen genes including LAMB4 were confirmed with prognostic value in USs; immune infiltration analysis revealed the prognositic value of myeloid dendritic cells, plasmacytoid dendritic cells, natural killer cells, macrophage M1, monocytes and hematopoietic stem cells in USs; the deep learning model named Max-Mean Non-Local multi-instance learning (MMN-MIL) showed satisfactory performance in predicting the survival of US patients, with the area under the receiver operating curve curve reached 0.909 and accuracy achieved 0.804. Conclusion USs harbored distinct gene fusion characteristics and gene expression features between HGESS, LGESS, and uLMS. The MMN-MIL model could effectively predict the survival of US patients.

Objective:To compare the efficacy of cannulated screw internal fixation combined with quadratus femoris bone flap with preservation of the posterior superior retinaculum and cannulated screw internal fixation alone in the treatment of femoral neck fracture in young and middle-aged patients.Methods:A retrospective cohort study was conducted to analyze the clinical data of 83 young and middle-aged patients with femoral neck fracture admitted to the 920th Hospital of Joint Logistic Support Force of PLA from January 2018 to January 2023, including 56 males and 27 females, aged 28-55 years [(42.7±3.2)years]. According to Garden classification, the fractures were classified as type III in 22 patients and type IV in 61. Based on Pauwels classification, the fractures were classified as type I in 15 patients, type II in 38 and type III in 30. Forty patients were treated with cannulated screw internal fixation combined with modified quadratus femoris bone flap (cannulated screw combined with bone flap group) and 43 with cannulated screw internal fixation alone (cannulated screw group). The two groups were compared in terms of the operation time, intraoperative blood loss, time to weight-bearing, length of hospital stay, and wound healing. The visual analogue scale (VAS) scores and Harris hip function scores at 1, 3, 6, 12 months after surgery and at the last follow-up. The postoperative complication rate was detected.Results:All the patients were followed up for 20-70 months [(40.0±1.2)months]. The operation time and intraoperative blood loss were (105.2±2.7)minutes and (100.6±16.3)ml in the cannulated screw combined with bone flap group, which were longer or more than (92.4±4.7)minutes and (92.5±14.6)ml in the cannulated screw group ( P<0.01). The time to weight-bearing was (12.1±1.4)weeks in the cannulated screw combined with bone flap group, shorter than (23.6±1.2)weeks in the cannulated screw group ( P<0.01). There was no statistically significant difference in the length of hospital stay between the two groups (P>0.05). The incisions in both groups were healed by first intention. At 1 month after surgery, no statistically significant difference was observed in VAS scores between the two groups ( P>0.05); at 3, 6, 12 months after surgery and at the last follow-up, the VAS scores were (6.6±0.2)points, (4.5±0.3)points, (3.2±0.5)points, and (2.6±0.4)points in the cannulated screw combined with bone flap group, lower than (7.0±0.1)points, (5.2±0.2)points, (3.9±0.4)points, and (3.3±0.1)points in the cannulated screw group ( P<0.05 or 0.01). At 1 and 3 months after surgery, no statistically significant difference was observed in the Harris hip function scores between the two groups ( P>0.05); at 6, 12 months after surgery and at the last follow-up, the Harris hip function scores were (82.2±1.7)points, (90.0±1.4)points, and (91.6±1.0)points in the cannulated screw combined with bone flap group, higher than (75.2±1.7)points, (83.4±1.9)points, and (85.2±0.7)points in the cannulated screw group ( P<0.01). At the last follow-up, in the cannulated screw combined with bone flap group, the Harris hip function was rated excellent in 32 patients, good in 5, and fair in 3, with an excellent and good rate of 92.5%, while in the cannulated screw group, the Harris hip function was rated excellent in 20 patients, good in 13, and fair in 10, with an excellent and good rate of 76.7% ( P<0.05). The postoperative complication rate was 5.0% (2/40) in the cannulated screw combined with bone flap group, significantly lower than 23.2% (10/43) in the cannulated screw group ( P<0.05). Conclusion:Compared with cannulated screw internal fixation alone, cannulated screw internal fixation combined with quadratus femoris bone flap with preservation of the posterior superior retinaculum has the advantages of earlier weight-bearing, less pain, better recovery of hip joint function, and lower incidence of postoperative complications in the treatment of femoral neck fracture in young and middle-aged patients, despite longer operation time and more intraoperative blood loss.

Salmonella has demonstrated considerable potential as a vaccine vector,exhibiting robust immunogenicity,ease of oral administration,and cost-effective production.Live attenuated Salmo-nella carrying heterologous antigens can induce both localized mucosal immunity and systemic a-daptive immune responses in hosts after successfully reaching the intestinal tract via oral delivery.Recent advances such as permanent deletion of virulence genes,regulated-delayed attenuation and lysis systems,have initially achieved a balance between the safety and immunogenicity of these vaccine platforms.Nevertheless,practical applications of such vaccine vectors remain constrained by challenges related to gastrointestinal barrier obstruction,inefficient antigen delivery,and im-mune tolerance.With the rapid advancement of multidisciplinary technologies,these limitations are anticipated to be progressively addressed.This review presented a comprehensive summary and dis-cussion of the immune mechanisms,development strategies,current applications,advantages,and challenges associated with oral live attenuated Salmonella vaccine vectors.It also delineated the fu-ture direction of research and development,with a view to providing theoretical references for re-lated research.

Objective:To investigate the predictive value of computed tomography(CT) based radiomics model for the prognosis of patients with pancreatic ductal adenocarcinoma(PDAC).Methods:The retrospective cohort study was conducted. The clinicopathological data of 206 PDAC patients who were admitted to Zhongshan Hospital of Fudan University from August 2018 to December 2020 were collected. There were 115 males and 91 females, aged (64±9)years. All 206 pati-ents underwent enhanced CT examination. Based on radom number table, the 206 patients were randomly divided into a training set of 165 cases and a validation set of 41 cases with a ratio of 4∶1. The training set was used to construct the prediction model, and the test set was used to validate the performance of the prediction model. Observation indicators: (1) follow-up; (2) analysis of prognostic factors of PDAC patients in the training set; (3) construction and evaluation of prediction model for prognosis of PDAC patients. Comparison of measurement data with normal distribution between groups was conducted using the t test. Comparison of measurement data with skewed distribution between groups was conducted using the Wilcoxon W test. Comparison of count data between groups was conducted using the chi-square test or corrected chi-square test. The Kaplan-Meier method was used to calculate the survival rate and Log-rank test was used for survival analysis. Univariate and multivariate analyses were conducted using the COX regression model. The PyCharm software was used for the least absolute shrinkage and selection operator method (LASSO)-COX regression analysis. The receiver operating characteristic curve was plotted to evaluate the performance of radiomics model. Results:(1)Follow-up. Of the 206 patients,205 cases were followed up for 17.1(range, 12.0?40.1)months. The postoperative 1-, 2-, 3-year survival rates were 80.10%, 29.61% and 4.85%. (2) Analysis of prognostic factors for PDAC patients in the training dataset. Results of multivariate analysis showed that pathological N stage was an independent influencing factor for prognosis of PDAC patients in the training set ( hazard ratio=1.476, 95% confidence interval as 1.054?2.067, P<0.05). (3) Construction and evaluation of prediction model for prognosis of PDAC patients. A total of 1 595 radiomics features were finally extracted from the 206 patients. By intra-group feature selection and dimensionality reduction using LASSO-COX regression model, 10 radiomics features were obtained. Combined with 10 radiomics features and 11 clinical features, using the LASSO-COX regression analysis, 15 features were finally extracted to construct the CT based radiomics model for predicting prognosis of PDAC. The areas under receiver operating characteristic curve of the prediction model in predicting 2-year and 3-year overall survival rates of PDAC patients in the training set were 0.834 (95% confidence interval as 0.777?0.891) and 0.883 (95% confidence interval as 0.834?0.932), respectively. The area under curve of the prediction model for patients in the validation set was 0.606 (95% confidence interval as 0.456?0.756) and 0.625 (95% confidence interval as 0.477?0.773). Conclusion:The prediction model constructed on CT based radiomics features and clinical features for predicting the prognosis of PDAC patients shows a promising prediction efficiency.

Salmonella has demonstrated considerable potential as a vaccine vector,exhibiting robust immunogenicity,ease of oral administration,and cost-effective production.Live attenuated Salmo-nella carrying heterologous antigens can induce both localized mucosal immunity and systemic a-daptive immune responses in hosts after successfully reaching the intestinal tract via oral delivery.Recent advances such as permanent deletion of virulence genes,regulated-delayed attenuation and lysis systems,have initially achieved a balance between the safety and immunogenicity of these vaccine platforms.Nevertheless,practical applications of such vaccine vectors remain constrained by challenges related to gastrointestinal barrier obstruction,inefficient antigen delivery,and im-mune tolerance.With the rapid advancement of multidisciplinary technologies,these limitations are anticipated to be progressively addressed.This review presented a comprehensive summary and dis-cussion of the immune mechanisms,development strategies,current applications,advantages,and challenges associated with oral live attenuated Salmonella vaccine vectors.It also delineated the fu-ture direction of research and development,with a view to providing theoretical references for re-lated research.

Objective:To investigate the predictive value of computed tomography(CT) based radiomics model for the prognosis of patients with pancreatic ductal adenocarcinoma(PDAC).Methods:The retrospective cohort study was conducted. The clinicopathological data of 206 PDAC patients who were admitted to Zhongshan Hospital of Fudan University from August 2018 to December 2020 were collected. There were 115 males and 91 females, aged (64±9)years. All 206 pati-ents underwent enhanced CT examination. Based on radom number table, the 206 patients were randomly divided into a training set of 165 cases and a validation set of 41 cases with a ratio of 4∶1. The training set was used to construct the prediction model, and the test set was used to validate the performance of the prediction model. Observation indicators: (1) follow-up; (2) analysis of prognostic factors of PDAC patients in the training set; (3) construction and evaluation of prediction model for prognosis of PDAC patients. Comparison of measurement data with normal distribution between groups was conducted using the t test. Comparison of measurement data with skewed distribution between groups was conducted using the Wilcoxon W test. Comparison of count data between groups was conducted using the chi-square test or corrected chi-square test. The Kaplan-Meier method was used to calculate the survival rate and Log-rank test was used for survival analysis. Univariate and multivariate analyses were conducted using the COX regression model. The PyCharm software was used for the least absolute shrinkage and selection operator method (LASSO)-COX regression analysis. The receiver operating characteristic curve was plotted to evaluate the performance of radiomics model. Results:(1)Follow-up. Of the 206 patients,205 cases were followed up for 17.1(range, 12.0?40.1)months. The postoperative 1-, 2-, 3-year survival rates were 80.10%, 29.61% and 4.85%. (2) Analysis of prognostic factors for PDAC patients in the training dataset. Results of multivariate analysis showed that pathological N stage was an independent influencing factor for prognosis of PDAC patients in the training set ( hazard ratio=1.476, 95% confidence interval as 1.054?2.067, P<0.05). (3) Construction and evaluation of prediction model for prognosis of PDAC patients. A total of 1 595 radiomics features were finally extracted from the 206 patients. By intra-group feature selection and dimensionality reduction using LASSO-COX regression model, 10 radiomics features were obtained. Combined with 10 radiomics features and 11 clinical features, using the LASSO-COX regression analysis, 15 features were finally extracted to construct the CT based radiomics model for predicting prognosis of PDAC. The areas under receiver operating characteristic curve of the prediction model in predicting 2-year and 3-year overall survival rates of PDAC patients in the training set were 0.834 (95% confidence interval as 0.777?0.891) and 0.883 (95% confidence interval as 0.834?0.932), respectively. The area under curve of the prediction model for patients in the validation set was 0.606 (95% confidence interval as 0.456?0.756) and 0.625 (95% confidence interval as 0.477?0.773). Conclusion:The prediction model constructed on CT based radiomics features and clinical features for predicting the prognosis of PDAC patients shows a promising prediction efficiency.

Objective:To compare the efficacy of cannulated screw internal fixation combined with quadratus femoris bone flap with preservation of the posterior superior retinaculum and cannulated screw internal fixation alone in the treatment of femoral neck fracture in young and middle-aged patients.Methods:A retrospective cohort study was conducted to analyze the clinical data of 83 young and middle-aged patients with femoral neck fracture admitted to the 920th Hospital of Joint Logistic Support Force of PLA from January 2018 to January 2023, including 56 males and 27 females, aged 28-55 years [(42.7±3.2)years]. According to Garden classification, the fractures were classified as type III in 22 patients and type IV in 61. Based on Pauwels classification, the fractures were classified as type I in 15 patients, type II in 38 and type III in 30. Forty patients were treated with cannulated screw internal fixation combined with modified quadratus femoris bone flap (cannulated screw combined with bone flap group) and 43 with cannulated screw internal fixation alone (cannulated screw group). The two groups were compared in terms of the operation time, intraoperative blood loss, time to weight-bearing, length of hospital stay, and wound healing. The visual analogue scale (VAS) scores and Harris hip function scores at 1, 3, 6, 12 months after surgery and at the last follow-up. The postoperative complication rate was detected.Results:All the patients were followed up for 20-70 months [(40.0±1.2)months]. The operation time and intraoperative blood loss were (105.2±2.7)minutes and (100.6±16.3)ml in the cannulated screw combined with bone flap group, which were longer or more than (92.4±4.7)minutes and (92.5±14.6)ml in the cannulated screw group ( P<0.01). The time to weight-bearing was (12.1±1.4)weeks in the cannulated screw combined with bone flap group, shorter than (23.6±1.2)weeks in the cannulated screw group ( P<0.01). There was no statistically significant difference in the length of hospital stay between the two groups (P>0.05). The incisions in both groups were healed by first intention. At 1 month after surgery, no statistically significant difference was observed in VAS scores between the two groups ( P>0.05); at 3, 6, 12 months after surgery and at the last follow-up, the VAS scores were (6.6±0.2)points, (4.5±0.3)points, (3.2±0.5)points, and (2.6±0.4)points in the cannulated screw combined with bone flap group, lower than (7.0±0.1)points, (5.2±0.2)points, (3.9±0.4)points, and (3.3±0.1)points in the cannulated screw group ( P<0.05 or 0.01). At 1 and 3 months after surgery, no statistically significant difference was observed in the Harris hip function scores between the two groups ( P>0.05); at 6, 12 months after surgery and at the last follow-up, the Harris hip function scores were (82.2±1.7)points, (90.0±1.4)points, and (91.6±1.0)points in the cannulated screw combined with bone flap group, higher than (75.2±1.7)points, (83.4±1.9)points, and (85.2±0.7)points in the cannulated screw group ( P<0.01). At the last follow-up, in the cannulated screw combined with bone flap group, the Harris hip function was rated excellent in 32 patients, good in 5, and fair in 3, with an excellent and good rate of 92.5%, while in the cannulated screw group, the Harris hip function was rated excellent in 20 patients, good in 13, and fair in 10, with an excellent and good rate of 76.7% ( P<0.05). The postoperative complication rate was 5.0% (2/40) in the cannulated screw combined with bone flap group, significantly lower than 23.2% (10/43) in the cannulated screw group ( P<0.05). Conclusion:Compared with cannulated screw internal fixation alone, cannulated screw internal fixation combined with quadratus femoris bone flap with preservation of the posterior superior retinaculum has the advantages of earlier weight-bearing, less pain, better recovery of hip joint function, and lower incidence of postoperative complications in the treatment of femoral neck fracture in young and middle-aged patients, despite longer operation time and more intraoperative blood loss.